CN108761073A - A kind of mycoplasma pneumoniae antigen detection kit and preparation method thereof - Google Patents
A kind of mycoplasma pneumoniae antigen detection kit and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to technical field of in vitro diagnostic reagents, and in particular to a kind of mycoplasma pneumoniae antigen detection kit, including box body, box body is interior to be equipped with detection card and sample extraction liquid bottle；Detection card includes Ka Gai, card body and test strips, test strips are equipped with sample bed course, colloid layer gold, nitrocellulose filter and water accepting layer successively, sample pad solid phase biological element marks anti-mycoplasma pneumoniae monoclonal antibody A, the anti-mycoplasma pneumoniae monoclonal antibody B of colloid layer gold solid phase；Nitrocellulose filter is equipped with the nature controlling line of the detection line and coating sheep anti-mouse igg antibody of coating Streptavidin successively.The invention further relates to the preparation methods of the kit；Detection kit high sensitivity in the present invention can detect the mycoplasma pneumoniae antigen in throat swab sample.
Description
Technical field
The present invention relates to vitro diagnostic techniques fields, and in particular to a kind of mycoplasma pneumoniae antigen detection kit and its system
Preparation Method.
Background technology
Mycoplasma pneumoniae (Mycoplasma pneumonia) can be lived on one's own life most between thin CFU and virus
Small microorganism, size 200nm.It is caused to distribute the respiratory tract infection with small prevalence by mouth, nasal discharge air-borne transmission,
It is mainly seen in Children and teenager, has now found that also non-rare in adult, autumn and winter is more.Eaton agent pneumonia is that pneumonia branch is former
Acute respiratory infection caused by body accounts for about the 10% of 1/3 or more or various pneumonia of non-thin CFU pneumonia, seriously with pneumonia
Eaton agent pneumonia may also lead to death.
Due to the difficulty of Culture Mycoplasma, the clinically detection of common mycoplasma pneumoniae is mainly Serologic detection.People
In primary infection mycoplasma pneumoniae, the IgM antibody and IgG antibody that can successively generate, since antibody continues to exist in vivo, because
This Serologic detection is not suitable for early diagnosing, and can be used as retrospective diagnosis.
It cannot achieve the purpose that early diagnosis with the method for being separately cultured and detecting antibody.And antigen detection can be accomplished morning
Phase quick diagnosis detects the mycoplasma pneumoniae antigen in phlegm, throat swab, bronchial perfusate.Current used method has
ELISA, polymerase chain reaction (PCR) method.But it is due to limited conditions, domestic not popularize also at present.Especially PCR methods are due to existing
Reagent problem, pollution problem, therefore had to cautiously in result judgement.It is wiped since mycoplasma pneumoniae antigen exists only in pharynx
In son, sputum, bronchus, these sample interfering substances are more, and mycoplasma pneumoniae content is relatively low, and conventional colloidal gold method is difficult inspection
It measures.In view of the problems of the existing technology the present invention, is provided a kind of mycoplasma pneumoniae antigen detection kit, is carried using sample
It takes liquid to separate interfering substance and extracts mycoplasma pneumoniae active ingredient, using biotin-avidin amplification system significantly to carry
High reagent sensitivity, to detect the mycoplasma pneumoniae antigen in throat swab sample.
Invention content
It is an object of the invention to be：In view of the deficienciess of the prior art, providing a kind of mycoplasma pneumoniae antigen detection
Kit, the detection kit high sensitivity can detect the mycoplasma pneumoniae antigen in throat swab sample.
To achieve the goals above, the technical scheme is that：
A kind of mycoplasma pneumoniae antigen detection kit, including box body, the box body is interior to be equipped with detection card and sample extraction
Liquid bottle；The detection card includes Ka Gai, card body and test strips, and the test strips are equipped with sample bed course, colloid layer gold, nitric acid successively
Cellulose membrane and water accepting layer；The sample bed course solid phase biological element marks anti-mycoplasma pneumoniae monoclonal antibody A, the colloidal gold
The anti-mycoplasma pneumoniae monoclonal antibody B of layer solid phase；The nitrocellulose filter is equipped with the detection line of coating Streptavidin successively
With the nature controlling line of coating sheep anti-mouse igg antibody.
As a kind of perferred technical scheme, the extraction solution in the sample extraction liquid bottle is by bovine serum albumin(BSA), folded
Nitrogen sodium and triton x-100 are formulated.
As an improvement technical solution, per 100mL extraction solution in the bovine serum albumin(BSA) containing 0.5~1g/mL
1~3mL of Sodium azide of 10~15mL and 0.03~0.05g/mL.
It is a further object to provide a kind of systems of the higher mycoplasma pneumoniae antigen detection kit of sensitivity
Preparation Method.
In order to realize the purpose of foregoing invention, the technical scheme is that：
A kind of preparation method of mycoplasma pneumoniae antigen detection kit, the preparation method comprises the following steps：
(1) preparation of sample extraction liquid
1~3mL of Sodium azide of the bovine serum albumin(BSA) 10~15mL and 0.03~0.05g/mL of 0.5~1g/mL are taken, is used
0.3~0.5g/mL triton x-100s are settled to 100mL, obtain sample extracting solution；
(2) preparation of colloidal gold and label
0.5~1g/mL aqueous solution of chloraurate 100mL, are heated to boiling, and 8~10g/mL trisodium citrate 1.5mL are added,
Mixing continues to boil 10~15 minutes, adjusts pH, is added with stirring the anti-mycoplasma pneumoniae monoclonal antibody B of 2~3mg, continues to stir
10~15 minutes, 8~10g/mL NaCl are added, shake up, centrifuging the sediment that 10~15min is obtained with 10000r/min is
Colloidal gold labeled monoclonal antibody；
(3) label of biotin
Take 1mg/mL N hydroxysuccinimides esters (BNHS)-dimethyl sulphoxide solution, be added 1~2mL, 1.2~
The anti-mycoplasma pneumoniae monoclonal antibody A of 1.5mg/mL persistently stir 2~3h at room temperature, abundant with buffer solution under the conditions of 4 DEG C
Sodium azide and BSA are added after dialysis, obtained biotinylated derivative sets 4 DEG C, is kept in dark place；
(4) immobilization of colloidal gold and biotinylated derivative
4.1 pretreatment：By the glass fibre 0.1mol of the BSA Han 0.5~1g/L and the PBS buffer solution of pH7.2~7.5
Processing, dries；
4.2 immobilization colloid gold label objects：It takes glass fibre to be put into stainless steel disc, measures colloid gold label object 30 after redissolving
~40mL pours into stainless steel disc, and gold solution substantially uniformity is made to absorb, dry, obtains gold-labelled pad；
4.3 immobilization biotinylated derivatives：Glass fibre is put into stainless steel disc, measures biotinylated derivative 30 after redissolving
~40mL pours into stainless steel disc, and gold solution substantially uniformity is made to absorb, dry, obtains sample pad；
(5) it is coated with nitrocellulose filter
5.1 coating Streptavidins：Streptavidin is diluted to 0.8 with the PBS buffer solution of pH7.2~7.5~
1.2mg/mL is sprayed on nitrocellulose filter, forms Streptavidin detection line；
5.2 coating sheep anti-mouse igg antibodies：Sheep anti-mouse igg antibody is diluted to 1.4 with the PBS buffer solution of pH7.2~7.5
~1.6mg/mL is sprayed on nitrocellulose filter made from above-mentioned steps 5.1, forms sheep anti-mouse igg antibody nature controlling line；
5.3 dry nitrocellulose filter made from above-mentioned steps 5.2 1.5~2 hours；
(6) preparation of test strips
Nitrocellulose filter prepared by above-mentioned steps 5.3 is pasted in the centre position of PVC board, it is fine to fix nitric acid in PVC board
Paste blotting paper in the top of the plain film location of dimension；The lower section of nitrocellulose film location is fixed in PVC board, prepared by gluing steps (4)
Gold-labelled pad and sample pad, after flattening, be cut into the test strips of 4.0mm wide；
(7) detection card installation
The test strips that step (6) is cut are placed in the groove of card body, Ka Gai and card body are fastened, mycoplasma pneumoniae is obtained
Antigen detection card；The sample extraction liquid that step (1) is obtained is packed into bottle, is respectively placed in box body with detection card, is obtained pneumonia
Mycoplasma antigen detection kit.
As an improvement technical solution, with the K of 0.1~0.2mol/L in the step (2)2CO3Or Na2CO3It adjusts
PH is 7-8.
As an improvement technical solution, dialysed with the phosphate buffers of pH7.2~7.5 in the step (3).
As an improvement technical solution, the end of the final concentration 0.3-0.5g/L, BSA of Sodium azide in the step (3)
A concentration of 0.8-1.2g/L.
As an improvement technical solution, colloid gold label object is pressed 1 with liquid is redissolved in the step 4.2：8-12's
Ratio is redissolved, and the redissolution liquid contains 0.5-1g/mL PEG20000,0.5-1g/mL trehaloses and 1-2g/mL Tween20.
As an improvement technical solution, biotinylated derivative is pressed 1 with liquid is redissolved in the step 4.3：8-12's
Ratio is redissolved, and the redissolution liquid contains 1-2g/mL PEG20000,0.5-1g/mL trehaloses and 1-2g/mL Tween20.
The present invention is had the following advantages compared with prior art using above technical scheme：
1, sample extraction liquid provided by the invention, bovine serum albumin(BSA) can protect destination protein active；Sodium azide can extend
The term of validity of sample extraction liquid；Triton x-100 can not only make antigen protein dissolve and separation, but also can retaining protein day
Right conformation greatly improves the extraction efficiency of mycoplasma pneumoniae；
2, in kit preparation process of the invention, with the K of 0.1~0.2mol/L2CO3Or Na2CO3Tune pH is 7-8,
Colloidal gold can be made preferably to be combined with anti-mycoplasma pneumoniae monoclonal antibody A, obtain stable colloid gold label object, it is raw simultaneously
The ratio of object element (BNHS) and anti-mycoplasma pneumoniae monoclonal antibody B are particularly significant, since antibody volume is big compared with biotin,
If amount of antibody is more, flocks together and will produce steric hindrance；The present invention is using the mixed of PEG20000, trehalose and Tween20
Redissolution liquid of the liquid as colloid gold label object and biotinylated derivative is closed, colloid gold label object and biotinylated derivative are more conducive to
Better solid phase is on the glass fibers；
3, the present invention utilizes " Avidin-Biotin amplification system ", significantly improves reagent detection sensitivity, examination of the invention
Agent box detection sensitivity is up to 103CFU/mL has preferable specificity, can detect pneumonia branch before antibody appearance
The case where mycoplasma antigen, accurate judgement positive infection, provides foundation for treatment；
4, the technical solution adopted in the present invention reduces many red tapes, and convenient, fast, intuitive, it is special to be not required to
Instrument is also not required to professional training, and operation can be completed in by specification, and overall process only needs 20 minutes, reduces use cost, is suitble to
The grass-roots units such as hospital.
Description of the drawings
Fig. 1 is the overlooking structure diagram of test strips in the embodiment of the present invention；
Fig. 2 is the side structure schematic view of test strips in the embodiment of the present invention；
Wherein, 1- test strips, 11- sample bed courses, 12- colloid layer gold, 13- nitrocellulose filters, 131- detection lines, 132-
Nature controlling line, 14- water accepting layers.
Specific implementation mode
In order to make the purpose , technical scheme and advantage of the present invention be clearer, the present invention is carried out further detailed
Explanation.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not intended to limit the present invention.
Embodiment 1
A kind of mycoplasma pneumoniae antigen detection kit, including box body and box cover, the interior detection card that is equipped with of box body are carried with sample
Liquid taking bottle, detection card includes Ka Gai, card body and test strips, as depicted in figs. 1 and 2, test strips 1 successively equipped with sample bed course 11,
Colloid layer gold 12, nitrocellulose filter 13 and water accepting layer 14, sample bed course solid phase have the anti-mycoplasma pneumoniae list of biotin labeling
Clonal antibody A；It is anti-mycoplasma pneumoniae monoclonal antibody B that colloid layer gold solid phase, which has colloid gold label object,；Nitrocellulose filter according to
The nature controlling line 132 of the secondary detection line 131 for being equipped with coating Streptavidin and coating sheep anti-mouse igg antibody.
Embodiment 2
A kind of preparation method of mycoplasma pneumoniae antigen detection kit, includes the following steps：
(1) preparation of sample extraction liquid
By the Sodium azide 1mL of the bovine serum albumin(BSA) 10mL and 0.05g/mL of 1g/mL, with 0.5%g/mL triton x-100s
It is settled to 100mL, is packed into reagent bottle, obtains sample extracting solution；
(2) preparation of colloidal gold and label
1g/mL aqueous solution of chloraurate 100mL, are heated to boiling, 10% trisodium citrate 1.5mL of addition, rapid mixing, after
It is continuous to boil 10 minutes, with 0.2mol/L K2CO3PH is adjusted, anti-mycoplasma pneumoniae monoclonal antibody B 2mg is added with stirring, continues to stir
It mixes 10 minutes, 10g/mL NaCl is added, shake up, be colloid gold label with the 10000r/min centrifugations 10min sediments obtained
Object；
(3) label of biotin
With 1mL dmso solution 1mg biotin N hydroxysuccinimide esters, 1mg/mL N maloyls are obtained
Imines ester-dimethyl sulphoxide solution, is added 2mL, and a concentration of anti-mycoplasma pneumoniae monoclonal antibody A of 1.5mg/mL are held at room temperature
Continuous stirring 3h is fully dialysed with pH7.4 phosphate buffers (PBS) under the conditions of 4 DEG C, removes free biotin, be added folded
Nitrogen sodium (final concentration 0.5g/L) and 1.0g/L BSA will set 4 DEG C in conjunction with product, be kept in dark place；
(4) immobilization of colloidal gold and biotinylated derivative
4.1 pretreatment：0.1mol pH7.4PBS buffer solution of the glass fibre containing 0.5%BSA is handled, is dried；
4.2 immobilization colloid gold label objects：Glass fibre is put into stainless steel disc, measures colloid gold label object after redissolving
34mL pours into stainless steel disc, and gold solution substantially uniformity is made to absorb, and push-in drying room is dried 12 hours；
4.3 immobilization biotinylated derivatives：Glass fibre is put into stainless steel disc, measures biotinylated derivative after redissolving
34mL pours into stainless steel disc, and gold solution substantially uniformity is made to absorb, and push-in drying room is dried 12 hours；
4.4 immobilization colloid gold label objects are gold-labelled pad, and immobilization biotinylated derivative is sample pad；
(5) it is coated with nitrocellulose filter
5.1 coating Streptavidins：With the PBS buffer solution of pH7.4 by Streptavidin to 1.0mg/mL, with Membrane jetter with
Pump speed 16mm/s, film speed 320mm/s are sprayed on nitrocellulose filter, form detection line；
5.2 coating sheep anti-mouse igg antibodies：Sheep anti-mouse igg antibody is diluted to 1.5mg/mL with the PBS buffer solution of pH7.4,
It is sprayed on nitrocellulose filter made from above-mentioned steps 5.1 with pump speed 16mm/s, film speed 320mm/s with Membrane jetter, forms Quality Control
Line；
Nitrocellulose filter made from above-mentioned steps 5.2 is set drying 2 hours in 37 DEG C of baking ovens by 5.3；
(6) preparation of test strips
6.1 paste nitrocellulose filter prepared by above-mentioned steps 5.3 in the centre position of PVC board；
6.2 fix the top of nitrocellulose film location in PVC board, paste blotting paper, and blotting paper covers nitrocellulose
1.0 millimeters of film edge；
6.3 fix the lower section of nitrocellulose film location in PVC board, colloid gold label object prepared by gluing steps (5)
Gold-labelled pad and the sample pad for indicating Biotin-Antibody conjugate, glass fibre gold-labelled pad cover 1.0 millimeters of nitrocellulose filter,
Polyester film sample pad covers 1.0 millimeters of gold-labelled pad；
After 6.4 flatten the product that above-mentioned steps 6.3 obtain, it is cut into the test strips of 4.0mm wide；
(7) test strips of above-mentioned preparation are placed in the groove of card body by card taking lid and card body, and Ka Gai and card body are fastened,
Obtain mycoplasma pneumoniae antigen detection card.
(8) the detection card that the sample extraction liquid and step (7) that detection card obtains after installing with step (1) obtain is set respectively
In in box body, obtaining mycoplasma pneumoniae antigen detection kit.
Embodiment 3
The application method of kit：
(1) it detects
A, 0.5mL sample extraction liquid is added in attached Reagent Tube.
B, the swab stick after collecting sample is immersed in the extracting solution in Reagent Tube and is stirred, from the outside of Reagent Tube, use finger
It squeezes swab stick for several times, so that extracting solution is thoroughly impregnated swab stick, then extract swab stick, the liquid twisted out is to be measured as sample.
C, 3 drop sample liquids (about 100 μ L) are added dropwise to the well of detection card, observation is as a result, result after twenty minutes within 15 minutes
In vain.
(2) result judges
A, feminine gender：Only there are a red stripes in quality control region (C), occurs in test section (T) interior redfree band.Feminine gender knot
Fruit shows：Range can be examined by being less than without mycoplasma pneumoniae antigen or content in sample.
B, the positive：Two band occur.One red stripes is located in test section (T), and another red stripes are located at matter
It controls area (C).Positive findings show：Contain mycoplasma pneumoniae antigen in sample.
C, invalid：Quality control region (C) does not occur red stripes, shows that operating process is incorrect or detection card has damaged.
Embodiment 4
The sensitivity test of kit of the present invention
By mycoplasma pneumoniae reference culture culture to 107CFU/mL, then gradient dilution is carried out, concentration is as shown in Table 1 below,
Colloidal gold method kit provided by the invention is respectively adopted to be detected with common colloidal gold method kit, it is as a result as follows：
1 mycoplasma pneumoniae antigen detection sensitivity of table compares
As can be seen from the above table, it is 10 in mycoplasma pneumoniae standard concentration4When CFU/mL, two kinds of detection methods point
Not Jian Ce the testing results of 100 samples can show the positive；But it is 10 in mycoplasma pneumoniae standard concentration3When CFU/mL,
The kit testing result of the present invention has 97 the positive occur, and the testing result of common colloidal gold method kit has 79 sun occur
Property, definition of numerical value of the positive rate more than 90% (sample size is more than 20) as kit sensitivity according to testing result is said
The kit detection sensitivity of the bright present invention is up to 103CFU/mL。
Embodiment 5
The specificity (anti-interference ability) of kit of the present invention is tested
In clinical examination, detection sample is often comprising the similar other pathogens of close or clinical symptoms with antigenic structure, choosing
Take clinical definite parainfluenza virus (mixing), tuberculosis, chlamydia pneumoniae, adenovirus, Respiratory Syncytial Virus(RSV), pneumonia streptococcus CFU,
Positive (mycoplasma pneumoniae antigen is negative) each 10 of the sample of influenza A virus, influenza virus B type, is examined with the kit of the present invention
Survey, be as a result still feminine gender, it was demonstrated that this kit specificity is good, not with parainfluenza virus (mixing), tuberculosis, chlamydia pneumoniae,
Adenovirus, Respiratory Syncytial Virus(RSV), pneumonia streptococcus CFU, influenza A virus, influenza virus B type generate cross reaction.
This patent is not limited to above-mentioned specific embodiment, those skilled in the art from above-mentioned design,
Without performing creative labour, made various transformation are all fallen within the protection domain of this patent.
Claims (8)
1. a kind of mycoplasma pneumoniae antigen detection kit, including box body, the box body is interior to be equipped with detection card and sample extraction liquid
Bottle；The detection card includes Ka Gai, card body and test strips, and it is fine that the test strips are equipped with sample bed course, colloid layer gold, nitric acid successively
The plain film of dimension and water accepting layer, it is characterised in that：The sample bed course solid phase biological element marks anti-mycoplasma pneumoniae monoclonal antibody A,
The anti-mycoplasma pneumoniae monoclonal antibody B of colloid layer gold solid phase；It is affine that the nitrocellulose filter is equipped with coating strepto- successively
The nature controlling line of the detection line and coating sheep anti-mouse igg antibody of element；Extraction solution in the sample extraction liquid bottle is pure by ox blood
Albumen, Sodium azide and triton x-100 are formulated.
2. a kind of mycoplasma pneumoniae antigen detection kit according to claim 1, it is characterised in that：It is extracted per 100mL
1~3mL of Sodium azide of bovine serum albumin(BSA) 10~15mL and 0.03~0.05g/mL containing 0.5~1g/mL in solution.
3. a kind of preparation method of mycoplasma pneumoniae antigen detection kit as claimed in claim 1 or 2, it is characterised in that described
Preparation method includes the following steps：
(1) preparation of sample extraction liquid
1~the 3mL of Sodium azide for taking the bovine serum albumin(BSA) 10~15mL and 0.03~0.05g/mL of 0.5~1g/mL, with 0.3~
0.5g/mL triton x-100s are settled to 100mL, obtain sample extracting solution；
(2) preparation of colloidal gold and label
0.5~1g/mL aqueous solution of chloraurate 100mL, are heated to boiling, addition 8~10g/mL trisodium citrate 1.5mL, mixing,
Continue to boil 10~15 minutes, adjust pH, be added with stirring the anti-mycoplasma pneumoniae monoclonal antibody B of 2~3mg, continue stirring 10~
15 minutes, 8~10g/mL NaCl are added, shake up, it is colloid to centrifuge the sediment that 10~15min is obtained with 10000r/min
Golden labelled antibody；
(3) label of biotin
1mg/mL N hydroxysuccinimides ester-dimethyl sulphoxide solution is taken, 1~2mL, the anti-pneumonia of 1.2~1.5mg/mL is added
Mycoplasma monoclonal antibody A, persistently 2~3h of stirring is added folded under the conditions of 4 DEG C after fully being dialysed with buffer solution at room temperature
Nitrogen sodium and BSA, obtained biotinylated derivative are set 4 DEG C, are kept in dark place；
(4) immobilization of colloidal gold and biotinylated derivative
4.1 pretreatment：Glass fibre is handled with the 0.1mol of the BSA Han 0.5~1g/L and the PBS buffer solution of pH7.2~7.5,
It dries；
4.2 immobilization colloid gold label objects：Glass fibre is put into stainless steel disc, measure colloid gold label object 30 after redissolving~
40mL pours into stainless steel disc, and gold solution substantially uniformity is made to absorb, dry, obtains gold-labelled pad；
4.3 immobilization biotinylated derivatives：Glass fibre is put into stainless steel disc, measure biotinylated derivative 30 after redissolving~
40mL pours into stainless steel disc, and gold solution substantially uniformity is made to absorb, dry, obtains sample pad；
(5) it is coated with nitrocellulose filter
5.1 coating Streptavidins：Streptavidin is diluted to 0.8~1.2mg/mL with the PBS buffer solution of pH7.2~7.5,
It is sprayed on nitrocellulose filter, forms Streptavidin detection line；
5.2 coating sheep anti-mouse igg antibodies：Sheep anti-mouse igg antibody is diluted to 1.4 with the PBS buffer solution of pH7.2~7.5~
1.6mg/mL is sprayed on nitrocellulose filter made from above-mentioned steps 5.1, forms sheep anti-mouse igg antibody nature controlling line；
5.3 dry nitrocellulose filter made from above-mentioned steps 5.2 1.5~2 hours；
(6) preparation of test strips
Nitrocellulose filter prepared by above-mentioned steps 5.3 is pasted in the centre position of PVC board, nitrocellulose is fixed in PVC board
Paste blotting paper in the top of film location；The lower section of nitrocellulose film location, gold prepared by gluing steps (4) are fixed in PVC board
Mark pad and sample pad, flatten and are cut into test strips；
(7) detection card installation
It is placed in what step (6) was cut in card body, Ka Gai and card body is fastened, obtain mycoplasma pneumoniae antigen detection card；It will step
Suddenly the sample extraction liquid that (1) obtains is packed into bottle, is respectively placed in box body with detection card, and the detection examination of mycoplasma pneumoniae antigen is obtained
Agent box.
4. the preparation method of mycoplasma pneumoniae antigen detection kit according to claim 3, it is characterised in that：The step
(2) with the K of 0.1~0.2mol/L in2CO3Or Na2CO3Tune pH is 7-8.
5. the preparation method of mycoplasma pneumoniae antigen detection kit according to claim 3, it is characterised in that：The step
(3) it is dialysed with the phosphate buffers of pH7.2~7.5 in.
6. the preparation method of mycoplasma pneumoniae antigen detection kit according to claim 3, it is characterised in that：The step
(3) the final concentration of 0.8-1.2g/L of the final concentration 0.3-0.5g/L, BSA of Sodium azide in.
7. the preparation method of mycoplasma pneumoniae antigen detection kit according to claim 3, it is characterised in that：The step
Colloid gold label object is pressed 1 with liquid is redissolved in 4.2:The ratio of 8-12 is redissolved, and the redissolution liquid contains 0.5-1g/mL
PEG20000,0.5-1g/mL trehalose and 1-2g/mL Tween20.
8. the preparation method of mycoplasma pneumoniae antigen detection kit according to claim 3, it is characterised in that：The step
Biotinylated derivative is pressed 1 with liquid is redissolved in 4.3:The ratio of 8-12 is redissolved, the redissolution liquid contain 1-2g/mL PEG20000,
0.5-1g/mL trehaloses and 1-2g/mL Tween20.
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| CN110286236A (en) * | 2019-07-25 | 2019-09-27 | 科尼格沃斯(无锡)医学科技有限公司 | The detection method of mycoplasma pneumoniae detection kit, preparation method and mycoplasma pneumoniae |
| CN113546162A (en) * | 2021-05-31 | 2021-10-26 | 江苏省农业科学院 | Mycoplasma vaccine and preparation method thereof |
| CN114397451A (en) * | 2022-01-25 | 2022-04-26 | 山东康华生物医疗科技股份有限公司 | Chlamydia pneumoniae antigen detection plate and detection box |
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| CN113546162A (en) * | 2021-05-31 | 2021-10-26 | 江苏省农业科学院 | Mycoplasma vaccine and preparation method thereof |
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